If I get a fresh tissue just after surgery. Could I freeze the tissue and conserve it for ulterior use? Or, do I have to use a protocol of conservation with IRPM & Antibiotics and make the culture in the same day?
May we assume you mean freezing in liquid nitrogen? Make sure some sample is minced into 1 mm blocks, then freeze according to standard liquid nitrogen protocols for cells, using DMSO as Alex suggests. I recommend multiple samples.
Far better is to try the primary culture immediately. My experience in primary outgrowth is that even an hour or two delay had less success-- process into small pieces as soon as possible and attach by drying to the coated plate in a tiny droplet of serum, gently add medium, and incubate. Floaters won't grow out.
You might be able to bring a sterile large ziploc plastic bag containing all of the sterile components needed into the surgical theater-- razor blade, wax sheet, culture dishes, serum vial, transfer pipettes-- and use it like a glove box, mincing and plating the pieces immediately, then as they are drying return to the culture lab for drying down the pieces under the laminar flow hood for a few minutes. Add medium and incubate as soon as the pieces are adhered.
You want to try a test run first, and certainly practice primary cultures on the same normal tissue before fooling with the valuable tissue from surgery-- there are tricks to deal with the fibroblast competition if that's a factor. This practice can also help you select the best medium formulation for the tissue, if that's not obvious from the literature.
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